Immediate release pharmaceutical formulation of 4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2h-phthalazin-1-one

ABSTRACT

The present invention relates to a pharmaceutical formulation comprising the drug 4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one in a solid dispersion with a matrix polymer that exhibits low hygroscopicity and high softening temperature, such as copovidone. The invention also relates to a daily pharmaceutical dose of the drug provided by such a formulation. In addition, the invention relates to the use of a matrix polymer that exhibits low hygroscopicity and high softening temperature in solid dispersion with 4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one for increasing the bioavailability of the drug.

This application is a continuation of U.S. application Ser. No.15/449,353 filed Mar. 3, 2017 which is a continuation of U.S.application Ser. No. 14/688,326, filed Apr. 16, 2016, which is acontinuation of U.S. application Ser. No. 13/911,151, filed Jun. 6,2013, which is a continuation of U.S. application Ser. No. 12/574,801,filed Oct. 7, 2009 (now U.S. Pat. No. 8,475,842, issued Jul. 2, 2013),which claims the benefit under 35 U.S.C. § 119(e) of U.S. ApplicationNo. 61/103,347 filed on Oct. 7, 2008.

Certain embodiments of the invention disclosed herein were made under ajoint Research Agreement between Abbott GMBH & Co. KG and AstraZeneca UKLtd.

The present invention relates to novel pharmaceutical compositions withimproved bioavailability and/or stability and/or drug loading, toprocesses for preparing these novel pharmaceutical compositions and totheir use in treating cancer, either as a sole agent or in combinationwith other therapies.

In particular, the present invention relates to a pharmaceuticalformulation comprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature. A particularly suitablematrix polymer being copovidone. The invention also relates to a dailypharmaceutical dose of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneprovided by such a formulation. In addition, the invention relates tothe use of copovidone in a solid dispersion composition with4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onefor increasing the bioavailability and/or stability of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one,or for treating cancer in a patient.

4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one(Compound 1), which has the following structure:

is disclosed and exemplified in International Patent ApplicationPublication No. WO 2004/080976, (compound 168). It is apoly(ADP-ribose)polymerase (PARP) inhibitor currently in clinical trialsfor treating cancers, such as breast and ovarian cancer.

According to WO2005/012524 and WO2005/053662, PARP inhibitor compounds,such as4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one,are particularly effective in treating cancers whose cells are defectivein homologous recombination (HR) dependent DNA double-stranded break(DSB) repair pathway. BRCA1 (NM_007295) and BRCA 2 (NM_000059)hereditary breast/ovarian cancer genes are just 2 out of many proteinsin the HR dependent DNA DSB repair pathway. Other members of the HRdependent DNA DSB repair pathway include: ATM (NM_000051), ATR(NM_001184), DSS1 (U41515), RPA 1 (NM_002945.2), RPA 2 (NM_00294.6), RPA3 (NM_002974.3), RPA 4 (NM_013347.1), Chk1 (NM_001274.2), Chk2 (096017GI:6685284), RAD51 (NM_002875), RAD51L1 (NM_002877), RAD51c (NM_002876),RAD51L3 (NM_002878), DMC1 (NM_007068), XRCC2 (NM_005431), XRCC3(NM_05432), RAD52 (NM_002879), RAD54L (NM_003579), RAD54B (NM_012415),RAD50 (NM_005732), MRE11A (NM_005590) and NBS1 (NM_002485). Thus, forexample, breast or ovarian cancers that are BRCA1+ and/or BRCA2+ couldbe much more susceptible to treatment with a PARP inhibitor compound,than cancers without a defective homologous recombination (HR) dependentDNA double-stranded break (DSB) repair pathway; potentially allowingeffective monotherapy treatment, and/or treatment at lower doses withconcomitant fewer or lesser side effects.

4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one(Compound 1) is a weakly acidic compound with a pKa of about 12.5(phthalazinone moiety). It is essentially neutral across thephysiological pH range. The aqueous equilibrium solubility of Compound 1was measured to be around 0.10 mg/mL across a range of aqueous buffers(pH 1-9); this solubility is increased to 0.12-0.20 mg/mL in real andsimulated gastrointestinal media with the highest solubility of 0.20mg/mL in the fed state simulated intestinal fluid (see Example 1.1).

Compound 1 was determined to be moderately permeable, compared to thehigh permeability marker propranolol, when investigated using a Caco-2cell line. The Caco-2 Papp value was 3.67×10⁻⁶ cm/sec, which equates toa human Peff value of 1.4×10⁴ cm/sec. Compound 1 is at the limits ofpoorly soluble in terms of drug formulation being a tentative class 4(at doses above 25 mg) within the Biopharmaceutical ClassificationSystem (BCS) based on these solubility and permeability values (seeExample 1).

Predictions of the bioavailability of Compound 1, made based onsolubility and permeability measurements, suggested that an immediaterelease (IR) tablet would be suitable for Compound 1. Indeed, compoundswith similar solubility, permeability and dose range have beensuccessfully formulated as IR tablets (E.g. see Kasim et al. “Molecularproperties of WHO essential drugs and provision of biopharmaceuticsclassification.” Molecular Pharmaceutics. 1(1):85-96, 2004). When testedin dogs however, the exposure following administration of a conventionalIR tablet was much lower than expected (see Example 6; FIG. 13).

The oral bioavailability of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneto a patient is dependant to a certain extent upon the dissolution rateand solubility of the drug in the GI tract. The bioavailability of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onefor a series of formulations can be assessed by determining the areaunder the curve (AUC) of a graph of plasma4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneconcentration v. time elapsed since administration of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.

The inventors were able to address the poor bioavailability of an IRtablet of Compound 1 by making a lipidic formulation (Gelucire™ 44-14),and this formulation has been used in Phase I and II clinical trials.However, at high drug loading (>10%), reduced exposure was seen with thelipidic formulation (see Example 6 and FIG. 30). A potential issue withthe gelucire lipidic formulation was thus only realised during doseescalation studies aimed at determining the maximum tolerated dose and,thus predicting the potential therapeutic dose. It was realized that ifthe therapeutic dose was 400 mg, a 10% drug loaded Gelucire™ 44-14formulation would have to be administered as 16 size 0 capsules. Notonly does this present with patient compliance issues, it would alsohave commercial implications, e.g. increase in manufacturing, packaging,and transportation costs, etc.

In the event that4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneis required in daily dosages greater than 50 mg or 100 mg, (indeeddosages as high as 400 mg twice daily are being tested in clinicaltrials), it would be desirable to to find a formulation of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2h-phthalazin-1-onewith increased bioavailability and one that would allow a sufficientdrug loading to be achieved so that it could be administered by means ofa manageable number of units (e.g. fewer than 4 per day).

Such increased bioavailability could be useful in enabling a reductionin the daily dose of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onerequired to achieve comparable biological exposure seen with aconventional formulation, e.g. a conventional IR tablet of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.

There is a desire, therefore, to find a formulation of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onewith improved bioavailability and drug loading relative to aconventional IR tablet formulation, ideally a formulation with a targetbioavailability of around 90% (relative to an intravenous solution), anda formulation that permits sufficient drug loading to reduce the numberof units that need to be taken at any one time, for example fewer than 4and ideally to one or two units.

The present invention aims to provide a formulation of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onethat minimises the size and/or number of tablets or capsules requiredfor the therapeutically effective dose, ideally to fewer than 4 units,preferably only one or two units.

In terms of the aim of increasing the therapeutic potential of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one,the inventors sought to increase the therapeutic potential by achievingan increase in the bioavailability of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a formulation that permitted sufficient high drug loading (e.g.greater than 10%). In distinct embodiments the drug loading will be atleast 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55% or 60%. It will beappreciated that the greater the drug loading the greater the likelihoodof instability, so although it may be feasible to generate a formulationwith a 60% drug loading it may be preferable to adopt a lower drugloading so as to maintain stability.

Of the various formulation approaches available, the inventorsdiscovered that solid dispersion formulations with particular types ofpolymer were a means of addressing one or more of the aims stated above.Furthermore, it was surprisingly found that the solid dispersionformulations of the invention increased the bioavailability of Compound1 compared to the lipidic gelucire formulation.

The inventors have now surprisingly found that the therapeutic potentialof4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onecan be increased by formulating4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature. The matrix polymercopovidone was found to be particularly suitable as it could be used inhot melt extrusion without the need of a plasticiser and it provides aproduct with acceptable stability, even at 30% drug loading in the finalproduct (e.g. tablet).

It would be further desirable to identify a suitable matrix polymer thatcould be formulated into a solid dispersion with the drug using any ofthe available solid dispersion techniques without the need foradditional surfactants/plasticisers as it would be appreciated that thepresence of certain extraneous excipients could compromise the stabilityCompound 1 (e.g. the ability to remain in amorphous form).

Thus, in one embodiment the solid dispersion formulation of theinvention does not comprise a surfactant/plasticiser.

According to a first aspect of the invention there is provided apharmaceutical formulation comprising an active agent in soliddispersion with a matrix polymer, wherein the active agent is4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a salt or solvate thereof, and the matrix polymer exhibits lowhygroscopicity and high softening temperature.

In one embodiment the active agent is present in the formulation instable amorphous form. Where the active agent is present in theformulation in stable amorphous form, the formulation may stabilise theactive agent in the formulation in the amorphous form and may reduceconversion or reversion to other forms.

In certain embodiments it will be desirable for the salt or solvate ofCompound 1 to be a pharmaceutically acceptable salt or solvate.

As used herein, by ‘polymer’ we mean a macromolecule composed ofrepeating structural units connected by covalent chemical bonds. Theterm encompasses linear and branched polymers, cyclic polymers such ascyclic oligosaccharides (including cyclodextrins), homopolymers andcopolymers, whether natural, synthetic or semi-synthetic in origin.

As used herein, the term ‘matrix polymer’ means a material that exhibitslow hygroscopicity and high softening temperature comprising a polymeror a blend of two or more polymers.

As used herein, by “low hygroscopicity” we mean having an equilibriumwater content <10% at 50% relative humidity, as determined by DynamicVapour Sorption (DVS), disclosed in Bergren, M. S. Int. J. Pharm103:103-114 (1994).

As used herein, by “high softening temperature” we mean that thematerial, in “as received” form (that is to say, without having beenexposed to high humidity) exhibits a glass transition temperature (Tg)or melting point (Tm) >100° C., as determined by Differential Scanningcalorimetry (DSC). The person of ordinary skill in the art willappreciate that Tg is a measurement appropriate for polymers that are inan amorphous state or form and Tm is a measurement that is appropriatefor polymers that are in a crystalline state or form.

Suitable matrix polymers for use in the invention include: copovidone,hypromellose phthalate (hydroxypropylmethylcellulose phthalate, HPMCP),hypromellose acetate succinate (hydroxypropylmethylcellulose acetatesuccinate, HPMCAS), -2-hydroxypropyl-β-cyclodextrin (HPBCD),hypromellose (hydroxypropylmethylcellulose, HPMC), polymethacrylates(poly(methacrylic acid, methyl methacrylate 1:1; poly(methacrylic acid,ethyl acrylate) 1:1), hydroxypropyl cellulose (HPC), and celluloseacetate phthalate (CAP).

Copovidone is a synthetic, linear, random copolymer ofN-vinyl-2-pyrrolidone (VP) and vinyl acetate (VA) with the chemicalformula (C₆H₉NO)_(m) (C₄H₆O₂)_(n) where the VA content is nominally 40%(but may vary, for example between 35-41%). The addition of vinylacetate to the vinylpyrrolidone polymer chain reduces hygroscopicity andglass transition temperature (Tg) of the polymer relative to Povidone(polyvinyl pyrrolidone, PVP homopolymer).

The K-value for copovidone is between 25 and 31, and since the K-valueis calculated from the kinematic viscosity of a 1% aqueous solution, itis related to the average molecular weight of the polymer. The averagemolecular weight (Mw) ranges from ˜24,000 to 30,000.

According to one aspect of the invention there is provided apharmaceutical formulation comprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with copovidone. In one embodiment thepharmaceutical formulation is one suitable for mucosal administration toa patient. A particular mucosal administration route is oral, e.g. atablet or capsule, and the like.

The invention also provides a daily pharmaceutical dose of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onewherein the dose comprises a therapeutically effective amount of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature. In one embodiment thematrix polymer is copovidone. In a further embodiment the pharmaceuticalformulation is mucosally administrable to a patient.

In a particular embodiment, the therapeutically effective amount of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneis in the range 10 to 1000 mg, in a further embodiment the dosecomprises 25 to 400 mg of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.

According to a further aspect of the invention there is provided apharmaceutical formulation comprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with copovidone, and comprising one or moreadditional compounds useful in the treatment of cancer. In oneembodiment the pharmaceutical formulation is for mucosal administrationto a patient.

According to a further aspect of the invention there is provided an oralpharmaceutical composition comprising a solid amorphous dispersioncomprising an active agent and at least one matrix polymer, wherein thematrix polymer exhibits low hygroscopicity and high softeningtemperature and wherein the active agent is4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof.

Further aspects of the invention relate to the use of a matrix polymerthat exhibits low hygroscopicity and high softening temperature, such ascopovidone, in solid dispersion with4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof, in themanufacture of a medicament, particularly for treating cancer; and, amethod of treating cancer comprising administration to a patient in needthereof of a therapeutically effective amount of a formulationcomprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof, in soliddispersion with a matrix polymer that exhibits low hygroscopicity andhigh softening temperature, such as copovidone. In such aspects, themedicament may comprise from 10 to 1500 mg of Compound 1, such as from10 to 1000 mg and from 25-400 mg.

Further aspects of the invention relate to: a method for increasing thebioavailability of the drug4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a patient in need of said drug, comprising administering to saidpatient a formulation comprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature; and, a dailypharmaceutical dose of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4fluoro-benzyl]2H-phthalazin-1-one for treating cancer in the patient,wherein the dose comprises 10 to 1000 mg of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature. In a particularembodiment of these aspects the matrix polymer is copovidone.

According to a further aspect of the invention there is provided amethod of producing a solid amorphous dispersion of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onecomprising:

-   -   (i) mixing a suitable amount of        4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one        or a pharmaceutically acceptable salt or solvate thereof with a        desired amount of at least one matrix polymer, wherein the        matrix polymer exhibits low hygroscopicity and high softening        temperature;    -   (ii) increasing the temperature of the mixture to produce a        melt; and    -   (iii) extruding the melt to produce a solid product.

In step (iii) the melt may be extruded as a solid rod which may then befurther processed, for example by milling, to produce a powder suitablefor use in a pharmaceutical formulation. Alternatively, the melt may beextruded into one or more moulds. Such moulds may, for example providefor shaped products such as elliptical or tablet shapes.

In step (ii) the melt could be produced by applying thermal heat and/ormechanical stress.

According to the various aspects of the invention a particular ratio of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one:matrix polymer by weight is from 1:0.25 to 1:10. More preferably thelower limit of the range is 1:≥4, 1:5 or 1:7. Preferably, the upperlimit of this range is 1:≤2, 1:1, 1:0.5 or 1:0.3. Suitable ratios are1:2, 1:3 and 1:4. In one embodiment, the range is 1:≥2 to 1:10. Inanother embodiment, the solid dispersion includes a surface-active agentand/or a plasticiser. Further discussion of surface-active agents andplasticisers appears below.

As used herein, the phrase “therapeutically effective amount” means thedrug dosage that provides the specific pharmacological response forwhich the drug is administered in a significant number of subjects inneed of such treatment. It is emphasized that a therapeuticallyeffective amount of a drug that is administered to a particular subjectin a particular instance will not always be effective in treating theconditions/diseases described herein, even though such dosage is deemedto be a therapeutically effective amount by those of skill in the art.By way of example, the therapeutically effective amount of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onecould be 25 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 400 mg,500 mg, 600 mg or 750 mg once or twice a day.

The solid dispersion formulations of the invention exhibit increasedbioavailability and drug loading potential and are thus likely torequire fewer dose units compared to conventional/immediate release4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneformulations.

One aspect of the invention provides a daily pharmaceutical dose of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onefor treating cancer in a patient, wherein the dose comprises 10 to 1500mg of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein a solid dispersion with a matrix polymer that exhibits lowhygroscopicity and high softening temperature, such as copovidone. Inone embodiment the pharmaceutical dose is administrable to a patientmucosally. In another embodiment the dose comprises 25 to 600 mg of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.

In various embodiments, the dose comprises 1500, 1250, 1000, 800, 700,600, 500, 450, 400, 300, 250, 225, 200, 175, 150, 125, 100, 75, 50, 25,15 or 10 mg of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.In particular embodiments, the dose comprises 25, 50, 100, 200 or 400 mgof4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one.

Additional excipients may be included in the formulation or dose. Forexample, the formulation or dose may comprise one or more fillers,binders, disintegrants and/or lubricants.

Suitable fillers include, for example, lactose, sugar, starches,modified starches, mannitol, sorbitol, inorganic salts, cellulosederivatives (e.g. microcrystalline cellulose, cellulose), calciumsulphate, xylitol and lactitol.

Suitable binders include, for example, lactose, starches, modifiedstarches, sugars, gum acacia, gum tragacanth, guar gum, pectin, waxbinders, microcrystalline cellulose, methylcellulose,carboxymethylcellulose, hydroxypropyl methylcellulose, hydroxyethylcellulose, hydroxypropyl cellulose, copolyvidone, gelatine,polyvinylpyrollidone (PVP) and sodium alginate.

Suitable disintegrants include, for example, crosscarmellose sodium,crospovidone, polyvinylpyrrolidone, sodium starch glycollate, cornstarch, microcrystalline cellulose, hydroxypropyl methylcellulose andhydroxypropyl cellulose.

Suitable lubricants include, for example, magnesium stearate, magnesiumlauryl stearate, sodium stearyl fumarate, stearic acid, calciumstearate, zinc stearate, potassium benzoate, sodium benzoate, myristicacid, palmitic acid, mineral oil, hydrogenated castor oil, medium-chaintriglycerides, poloxamer, polyethylene glycol and talc.

Additional conventional excipients, which may be added, includepreservatives, stabilisers, anti-oxidants, silica flow conditioners,antiadherents or glidants.

Other suitable fillers, binders, disintegrants, lubricants andadditional excipients which may be used are described in the Handbook ofPharmaceutical Excipients, 5th Edition (2006); The Theory and Practiceof Industrial Pharmacy, 3rd Edition 1986; Pharmaceutical Dosage Forms1998; Modern Pharmaceutics, 3rd Edition 1995; Remington's PharmaceuticalSciences 20th Edition 2000.

In certain embodiments, the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onewill be present in an amount of 10 to 70%, and preferably from 15 to 50%(more preferably 20 to 30% or 25 to 35%) by weight of the soliddispersion.

In certain embodiments, one or more fillers will be present in an amountof 1 to 70% by weight of the formulation or dose.

In certain embodiments, one or more binders will be present in an amountof 2 to 40% by weight of the formulation or dose.

In certain embodiments, one or more disintegrants will be present in anamount of 1 to 20%, and especially 4 to 10% by weight of the formulationor dose.

It will be appreciated that a particular excipient may act as both abinder and a filler, or as a binder, a filler and a disintegrant.Typically the combined amount of filler, binder and disintegrantcomprises, for example, 1 to 90% by weight of the formulation or dose.

In certain embodiments, one or more lubricants will be present in anamount of 0.5 to 3%, and especially 1 to 2% by weight of the formulationor dose.

In certain embodiments, one or more surface-active agents will bepresent in the solid dispersion in an amount of 0.1 to 50%, preferably≤5% (eg, 1 to 2%) by weight of the solid dispersion. The presence of asurface-active agent provides a further enhancement of the increase intherapeutic potential achieved with the present invention. Examples ofsuitable surface-active agents include: anionic surfactants such assodium dodecyl sulphate (sodium lauryl sulphate); docusate sodium;cationic surfactants such as cetrimide, benzethonium chloride,cetylpyridinium chloride and lauric acid; nonionic surfactants such aspolyoxyethylene alkyl ethers, polyoxyethylene sorbitan fatty acidesters, e.g. polysorbates 20, 40, 60 and 80; polyoxyethylene castor oilderivatives, e.g. Cremophor RH40™; polyoxyethylene stearates andpoloxamers.

In certain embodiments, one or more plasticisers will be present in thesolid dispersion in an amount of 0.1% to 50%, preferably ≤5% (e.g. 1 to2%) by weight of the solid dispersion. The presence of a plasticiser mayenhance processability of the solid dispersion, for example when a meltextrusion process is used. Examples of suitable plasticisers include:acetyltributyl citrate, acetyltriethyl citrate, benzyl benzoate,chlorbutanol, dextrin, dibutyl phthalate, diethyl phthalate, dimethylphthalate, glycerine, glycerine monostearate, mannitol, mineral oil,lanolin alcohols, palmitic acid, polyethylene glycol, polyvinyl acetatephthalate, propylene glycol, 2-pyrrolidone, sorbitol, stearic acid,triacetin, tributyl citrate, triethanolamine and triethyl citrate.

The term “solid dispersion” as used herein means systems in which anactive agent is dispersed in an excipient carrier. With respect to thestate of the drug in the systems, solid dispersions in this sense caninclude compositions in which the drug is dispersed as discrete domainsof crystalline or amorphous drug, or as individual molecules within anexcipient carrier. With respect to the complete drug-excipientcomposite, solid dispersions can be relatively large solid masses suchas pellets, tablets, films or strands; or they can exist as free flowingpowders consisting of micro- or nano-sized primary particles oraggregates thereof. The bulk state of the solid dispersion compositiondepends largely upon the mode of processing (Miller, D. A., McGinty, J.W., Williams III, R. O. Solid Dispersion Technologies.Microencapsulation of Oil-in-Water Emulsions 172 (2008) pp 451-491).

In the present invention the definition of a solid dispersion does notencompass physical mixtures from dry or wet mixing or dry blendingoperations.

Methods for preparing solid dispersions are known in the art andtypically comprise the steps of dissolving the drug and the polymer in acommon solvent and evaporating the solvent. The solvent can be routinelyselected according to the polymer used. Examples of solvents are:acetone, acetone/dichloromethane, methanol/dichloromethane,acetone/water, acetone/methanol, acetone/ethanol,dichloromethane/ethanol or ethanol/water. Methods for evaporatingsolvent include rotary evaporation, spray drying, lyophilisation andthin film evaporation. Alternatively solvent removal may be accomplishedby cryogenic freezing followed by lyophilisation. Other techniques maybe used such as melt extrusion, solvent controlled precipitation, pHcontrolled precipitation, supercritical fluid technology and cryogenicco milling.

This invention further discloses a method of making the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one:copovidone solid dispersion. Such a method comprises (i) dissolving asuitable amount of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneand matrix polymer in a common solvent; and (ii) removing the solvent.Pharmaceutical compositions comprising the dispersion can be made, forexample by adding such things as stabilizers and/or additionalexcipients as required. In a particular embodiment, the solvent isremoved by spray drying.

According to another aspect of the invention the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one:copovidonesolid dispersion is made by melt extrusion. Such a method comprisesadding the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one,or a pharmaceutically acceptable salt or solvate thereof, and copovidonepolymer, and any additional optional excipients, including plasticisers,to a melt extrusion apparatus which then heats and mixes and finallyextrudes the solid dispersion product. The extruder heats the mixture toa temperature high enough to melt the mixture but low enough so as tonot degrade the constituents.

According to another aspect of the invention there is provided a methodof producing a solid amorphous dispersion of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onecomprising simultaneously exposing4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof and at leastone matrix polymer, wherein the matrix polymer exhibits lowhygroscopicity and high softening temperature, to hot melt extrusion.

According to another aspect of the invention there is provided a methodof making a solid dispersion product of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2h-phthalazin-1-one,comprising:

(a) providing a powdered or granulated premix comprising:

(i) 5-60% by weight of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2h-phthalazin-1-one;and,

(ii) 40-95% copovidone;

(b) melting the premix, without addition of solvent, in a kneader or anextruder extruder to obtain a homogeneous melt, and(c) shaping and solidifying the melt to obtain a solid dispersionproduct.

In one embodiment, the solid dispersion product is formed into asuitable dosage form ready for oral administration.

In another embodiment, the solid dispersion product is ground up, mixedwith one or more additional excipients or ingredients, and tabletted orencapsulated into a suitable dosage form.

When referring to a solid dispersion we do not exclude the possibilitythat a proportion of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onemay be dissolved within the matrix polymer, the exact proportion, ifany, will depend upon the particular polymer selected.

In the formulations of the invention, at least some of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onemay be present in amorphous form in the solid dispersion with the matrixpolymer. The provision of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein amorphous form is additionally advantageous, since it furtherincreases the solubility and dissolution rate of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-one,thereby enhancing the increase in therapeutic potential achieved withthe present invention. Whether or not drug is present in amorphous formcan be determined by conventional thermal analysis or X-ray diffraction.In one embodiment, at least 25% of the4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein the formulation is present in amorphous form, as measured using XRPD.More preferably, this amount is at least 30%, 40%, 50%, 75%, 90%, 95%,as measured using XRPD. The most preferred embodiment is where 100% ofthe4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein the formulation is in amorphous form. In reality, current XRPD toolsand techniques may only be able to detect >5% crystalline form, and thusthe inability to detect crystalline form may mean that the sample isbetween 95% and 100% amorphous.

XRPD may be augmented by emerging nanometer-scale characterisationtechniques: Pair-wise Distribution Function (transformation of the X-raydiffraction pattern to a normalised scattering function) may facilitatethe detection of nanocrystallinity; Solid State NMR proton spindiffusion studies may be used to detect phase separation, as may AtomicForce Microscopy and Nanothermal analysis. Such techniques arecomparative rather than absolute but are useful tools in the developmentand optimisation of pharmaceutical solid dispersion formulations.

In a further embodiment, the drug is in stable amorphous form, by whichis meant that the stability (ability to remain in amorphous form andresist converting to crystalline form) of the amorphous state ofCompound 1 is extended in the solid dispersion formulation of theinvention relative to the stability of the amorphous state of Compound 1on its own.

In a preferred embodiment, the formulations and doses are mucosallyadministrable, i.e. administrable to mucosal membranes for absorptionacross the membranes. To this end, suitable routes of administrationinclude administration by inhalation, as well as oral, intranasal andrectal administration. Oral administration is particularly preferred. Atablet, capsule or other form of the formulation would be chosen by theskilled addressee according to the route of administration. Other routesof administration, e.g. parenteral are however not excluded.

The4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneis useful to provide a poly-ADP-ribose polymerase (PARP) inhibitoryeffect. This effect is useful for treating cancer, for example breast orovarian cancer, and particularly cancers that possess a defectivehomologous recombination (HR) dependent DNA double-stranded break (DSB)repair pathway, such as BRCA1+ and/or BRCA2+ve cancers.

Another aspect of the invention is directed to a4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onecomposition, comprising4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein solid dispersion with copovidone, and comprising one or moreadditional compounds useful in the treatment of cancer.

Particularly, useful “additional” anti-cancer compounds include DNAdamage promoting agents. A DNA damage promoting agent is a compound(such as a small organic molecule, peptide or nucleic acid) whichincreases the amount of DNA damage in a cell, either directly orindirectly, for example through inhibition of DNA repair. The DNA damagepromoting agent is often a small organic molecule compound.

Suitable DNA damage promoting agents include agents which damage DNA ina cell (i.e. DNA damaging agents), for example alkylating agents such asmethyl methanesulfonate (MMS), temozolomide, dacarbazine (DTIC),cisplatin, oxaliplatin, carboplatin,cisplatin-doxorubicin-cyclophosphamide, carboplatin-paclitaxel,cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan,etoposide, teniposide, amsacrine, irinotecan, topotecan and rubitecanand nitrosoureas, topoisomerase-1 inhibitors like Topotecan, Irinotecan,Rubitecan, Exatecan, Lurtotecan, Gimetecan, Diflomotecan(homocamptothecins); as well as 7-substituted non-silatecans; the7-silyl camptothecins, BNP 1350; and non-camptothecin topoisomerase-Iinhibitors such as indolocarbazoles, topoisomerase-II inhibitors likeDoxorubicin, Danorubicin, and other rubicins, the acridines (Amsacrine,m-AMSA), Mitoxantrone, Etopside, Teniposide and AQ4, dualtopoisomerase-I and II inhibitors like the benzophenazines, XR 11576/MLN576 and benzopyridoindoles, and antimetabolites such as gemcitabine,antifolates such as fluoropyrimidines like 5 fluorouracil and tegafur,raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea, andarsenic trioxide.

The patient can be a human, e.g. an adult or a child, but the treatmentof other mammals is also contemplated.

Aspects of the present invention will now be illustrated with referenceto the accompanying figures described below and experimentalexemplification, by way of example and not limitation. Further aspectsand embodiments will be apparent to those of ordinary skill in the art.

FIG. 1 shows permeability of Compound 1 across Caco-2 monolayers (n=3,±s.d.)

FIG. 2 shows in vitro dissolution of various Compound 1 formulations.

FIG. 3 shows a thermogram of a solid dispersion exhibiting a melttransition due to the presence of crystalline Compound 1

FIG. 4 shows an image of a tablet which exhibits a single crystal ofCompound 1 in the hot-stage microscopy method

FIG. 5 shows PDF spectra for solid dispersions of Compound 1 andcopovidone at various drug loadings

FIG. 6 shows a comparison of PDF spectra for solid dispersions ofCompound 1 and copovidone with simulated spectra for physical mixturesat various drug loadings

FIGS. 7(1)-7(6) show TM-AFM topographic (height), tip-deflection (error)and phase (mechanical property) images from 50 μm×50 μm and 10 μm×10 μmscans for solid dispersions of compound 1 and copovidone at 10% drugloading:—

FIG. 7(1) is the 50 μm×50 μm topographic (height)

FIG. 7(2) is the 50 μm×50 μm tip-deflection (error)

FIG. 7(3) is the 50 μm×50 μm phase (mechanical property)

FIG. 7(4) is the 10 μm×10 μm topographic (height)

FIG. 7(5) is the 10 μm×10 μm tip-deflection (error)

FIG. 7(6) is the 10 μm×10 μm phase (mechanical property)

FIGS. 8(1)-8(6) show TM-AFM topographic (height), tip-deflection (error)and phase (mechanical property) images from 50 μm×50 μm and 10 μm×10 μmscans for solid dispersions of compound 1 and copovidone at 30% drugloading:—

FIG. 8(1) is the 50 μm×50 μm topographic (height)

FIG. 8(2) is the 50 μm×50 μm tip-deflection (error)

FIG. 8(3) is the 50 μm×50 μm phase (mechanical property)

FIG. 8(4) is the 10 μm×10 μm topographic (height)

FIG. 8(5) is the 10 μm×10 μm tip-deflection (error)

FIG. 8(6) is the 10 μm×10 μm phase (mechanical property)

FIGS. 9(1)-9(6) show TM-AFM topographic (height), tip-deflection (error)and phase (mechanical property) images from 50 μm×50 μm and 10 μm×10 μmscans for solid dispersions of compound 1 and copovidone at 40% drugloading:—

FIG. 9(1) is the 50 μm×50 μm topographic (height)

FIG. 9(2) is the 50 μm×50 μm tip-deflection (error)

FIG. 9(3) is the 50 μm×50 μm phase (mechanical property)

FIG. 9(4) is the 10 μm×10 μm topographic (height)

FIG. 9(5) is the 10 μm×10 μm tip-deflection (error)

FIG. 9(6) is the 10 μm×10 μm phase (mechanical property)

FIG. 10 shows an XRPD diffractogram for Compound 1 Form H

FIG. 11 shows a representative DSC trace for Compound 1 Form H

FIG. 12 shows an XRPD diffractogram for Opadry

FIG. 13 shows an infrared spectrum of Compound 1

FIG. 14 shows infrared spectra of Aqoat MG, HP55S, Pharmacoat, Povidoneand Copovidone

FIG. 15 shows a synchronous spectrum of Aqoat MG annotated withcorrelation squares

FIG. 16 shows an asynchronous spectrum of Aqoat MG

FIG. 17 shows a synchronous spectrum of HP55S

FIG. 18 shows an asynchronous spectrum of HP55S

FIG. 19 shows an a synchronous spectrum of HP55S (high sensitivity)

FIG. 20 shows a synchronous spectrum of Pharmacoat

FIG. 21 shows an asynchronous spectrum of Pharmacoat

FIG. 22 shows an asynchronous spectrum of Pharmacoat (high sensitivity)

FIG. 23 shows a synchronous spectrum of Povidone

FIG. 24 shows a synchronous spectrum of Povidone (high sensitivity)

FIG. 25 shows an asynchronous spectrum of Povidone

FIG. 26 shows a synchronous spectrum of Copovidone

FIG. 27 shows a synchronous spectrum of Copovidone (high sensitivity)

FIG. 28 shows an asynchronous spectrum of Copovidone

FIG. 29 shows an asynchronous spectrum of Copovidone (high sensitivity)

FIG. 30 shows a plot of plasma concentration vs time for the variousCompound 1 formulations.

EXAMPLE 1. CHARACTERISTICS OF COMPOUND 1 1.1 Solubility

The solubility of crystalline Form A of Compound 1 was measured in waterand a range of pH buffered solutions representing the physiological pHrange. The physical form of any undissolved (or precipitated) Compound 1was not assessed by XRPD after solubility determination. Solubility dataare summarised in Table 1. The Form A crystalline form of Compound 1 isdisclosed in WO2008/047082.

TABLE 1 Solubility of crystalline Compound 1 (Form A) in a range ofbuffers representing the physiological pH range (mg · mL⁻¹) Media 1 hrpH 24 hr pH Water 0.124 5.6 0.109 6.0 0.1M HCl 0.128 1.2 0.114 1.2 pH 3Citrate Buffer 0.124 2.9 0.112 2.9 pH 6.8 Phosphate Buffer 0.111 6.90.096 6.9 pH 9 Buffer 0.116 8.9 0.102 8.8 0.1M NaOH 0.650 12.5 0.59912.4The solubility of Compound 1 was also measured in real and simulatedgastrointestinal media (Table 2). Solubility in HIF and FeSSIF wasnotably higher than buffer solubilities reported in Table 1.

TABLE 2 Solubility of crystalline Compound 1 (Form A) real and simulatedgastrointestinal media Equilibrium solubility Media (mg · mL⁻¹), 24 hrSimulated Gastric Fluid (SGF)¹ 0.12 Human Gastric Fluid (HGF)² 0.15 FedState Simulated Intestinal Fluid (FeSSIF)³ 0.2 Fasted State SimulatedIntestinal Fluid (FaSSIF)³ 0.13 Human Intestinal Fluid (HIF)² 0.17 ¹SGFcontains 3.2 g pepsin, 2.0 g sodium chloride, and 7.0 mL hydrochloricacid per litre. ²Pooled from healthy volunteers; supplied by UppsalaUniversitet, Box 256, 751 05 Uppsala, Sweden ³Marques, M. Dissolutionmedia simulating fasted and fed states. Dissolution Technologies (May2004) pp 16.

1.2 Permeability

Compound 1 was determined to be moderately permeable when compared tothe high permeability marker propranolol, investigated using a validatedCaco-2 cell line, results are summarised in Table 3 and FIG. 1. Compound1 was shown to have propensity for efflux by P-gp at low concentrations(10 μM), which was inhibited by the selective P-gp inhibitor Elacridar(GF120918; GG918;N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolyl)ethyl]phenyl)-9,10-dihydro-5-methoxy-9-oxo-4-acridinecarboxamide, hydrochloride salt.

TABLE 3 Permeability of Compound 1 across Caco-2 monolayers (n = 3,±S.D.), compared to the high permeability marker propranolol and theefflux marker digoxin P_(app) (cm · sec⁻¹) Concentation (μM) A-to-BB-to-A Efflux Ratio  10 3.67 ± 0.34 23.70 ± 2.84 6.5 10 with Elacridar10.34 ± 1.38  14.29 ± 0.93 1.4 260 7.75 ± 0.88 17.75 ± 1.19 2.3 700  8.4± 0.41 15.06 ± 1.42 1.8 Propranolol 19.97 ± 2.57  21.48 ± 0.33 1.1Digoxin 1.34 ± 0.03 12.22 ± 1.37 9.1 Key: A = apical; B = basolateral

See FIG. 1. EXAMPLE 2. POLYMER CHARACTERISTICS

TABLE 4 Characteristics of polymers used in pharmaceutical soliddispersion formulations Softening Point^(b) Hygroscopicity Tg Tm PolymerGrade Supplier (% w/w)^(a) (° C.) (° C.) Copovidone Kollidon VA64 BASFSE 5 106 N/A Povidone Kollidon 17PF 16 136 N/A Kollidon 25 155 N/AKollidon 30 168 N/A Hypromellose phthalate HP55S Shin-Etsu 4 145 N/A(HPMCP) HP55 Chemical Co., 145 N/A Hypromellose acetate Aqoat LF Ltd 4120 N/A succinate (HPMCAS) Aqoat LG 120 N/A Aqoat MG 130 N/A2-hydroxypropyl-β- Kleptose HP Roquette Freres 7 278 N/A cyclodextrin(HPBCD) Hypromellose (HPMC) Pharmacoat 606 Shin-Etsu 4 175 N/A ChemicalCo., Ltd Poly(methacrylic acid, Eudragit L100-55 Evonik Degussa 4 115N/A ethyl acrylate) 1:1 GmbH Poly(methacrylic acid, Eudragit L100 6 160^(#) N/A methyl methacrylate) 1:1 Poly(butylmethacrylate, EudragitE100 1  48 N/A (2-dimethylaminoethyl) methacrylate, methyl methacrylate)1:2:1 acid, ethyl acrylate) 1:1 Poly(methacrylic acid, Eudragit S100 11 160^(#) N/A methyl methacrylate) 1:2 Polyethylene glycol PEG 6000 FlukaAG 2 N/A 55-63 (PEG) Poloxamer Pluronic (Lutrol) BASF SE 2 N/A 52-57 F68Pluronic (Lutrol) N/A 52-57 F127 Hydroxypropyl cellulose Klucel EFHercules, Inc. 5 130 N/A (HPC) Cellulose acetate Aquacoat CPD FMC 6 176N/A phthalate (CAP) Biopolymer Key: N/A = Not Applicable ^(a)Equilibriumwater content at 50% Relative Humidity (literature values) ^(b)Softeningtemperature expressed as glass transition temperature (Tg) or meltingpoint (Tm) - suppliers data ^(#)Accurate determination not possible dueto chemical degradation

EXAMPLE 3. SCREENING STUDY—POLYMERIC DISPERSIONS 3.1 Protocol

TABLE 5 Protocol for the screening study of Compound 1 solid dispersions

^(a)Poloxamer F127, PVP K30, Hydroxypropyl cellulose, Copovidone andPolyacrylic acid were not assessed in DCM/MeOH ^(b)Only PVP K25, HPMCPhthalate and Kleptose were assessed without additive at 33% loading^(c)Kleptose/PVP K25 blend assessed using Acetone/MeOH solvent systemonly in ratios 5:70 and 10:65 at 25% drug loading and in ratios 5:45 and10:40 at 50% drug loading, without additive ^(d)Kleptose/HPMC606 blendassessed as described above for Kleptose/PVP K25 blend

3.2 Methodology

A series of 4% w/w solutions, comprising binary mixtures of Compound 1and each of the polymers in the proportions specified in the protocol,were prepared by weighing into 1.8 mL vials and dissolving in thespecified solvent system. Further solutions comprising ternary mixturesof Compound 1, polymer and surfactant were prepared in a similar manner.Solvent was removed by evaporation at 40° C. under nitrogen (10 mL/minflow, 0.7 bar pressure) for 15 minutes followed by drying overnightunder full vacuum to produce a solid dispersion.

The resulting samples were assessed using XRPD (Bruker GADDSdiffractometer; data collection at room temperature using CuKα radiationin the 2θ region between 1.5 and 41.5°), immediately after preparationand after storage for up to 1 month at 30° C. and 60% RH.

3.3 Results

TABLE 6 Results for the screening study of Compound 1 solid dispersionsXRPD (crystalline Compound 1) 30° C./60% RH Polymer Solvent System Drug(% w/w) Additive After Prep. 1 week 1 month PEG6000 DCM/MeOH 25 None N/DPresent N/T PEG6000 DCM/MeOH 50 None N/D Present N/T PEG6000Acetone/MeOH 25 None N/D Present N/T PEG6000 Acetone/MeOH 50 None N/DPresent N/T PEG6000 Acetone/MeOH 33 SLS N/D N/T Present PEG6000Acetone/MeOH 33 Tween 80 N/D N/T Present PEG6000 Acetone/MeOH 33 Doc. NaN/D N/T Present Poloxamer F68 DCM/MeOH 25 None N/D Present N/T PoloxamerF68 DCM/MeOH 50 None N/D Present N/T Poloxamer F68 Acetone/MeOH 25 NoneN/D Present N/T Poloxamer F68 Acetone/MeOH 50 None N/D N/D N/T PoloxamerF68 Acetone/MeOH 33 SLS N/D N/T Present Poloxamer F68 Acetone/MeOH 33Tween 80 N/D N/T Present Poloxamer F68 Acetone/MeOH 33 Doc. Na N/D N/TPresent Poloxamer F127 Acetone/MeOH 25 None N/D Present N/T PoloxamerF127 Acetone/MeOH 50 None N/D Present N/T PVP K25 DCM/MeOH 25 None N/DN/D N/T PVP K25 DCM/MeOH 50 None N/D N/D N/T PVP K25 Acetone/MeOH 25None Not harvested PVP K25 Acetone/MeOH 33 None N/D N/D N/T PVP K25Acetone/MeOH 50 None Not harvested PVP K25 Acetone/MeOH 33 SLS N/D N/TN/D PVP K25 Acetone/MeOH 33 Tween 80 N/D N/T N/D PVP K25 Acetone/MeOH 33Doc. Na N/D N/T N/D PVP K30 Acetone/MeOH 25 None N/D N/D N/T PVP K30Acetone/MeOH 50 None N/D N/D N/T HPMC-606 DCM/MeOH 25 None N/D N/D N/THPMC-606 DCM/MeOH 50 None N/D N/D N/T HPMC-606 Acetone/MeOH 25 None Notharvested HPMC-606 Acetone/MeOH 50 None Not harvested HPMC-606Acetone/MeOH 33 SLS N/D N/T N/D HPMC-606 Acetone/MeOH 33 Tween 80 N/DN/T N/D HPMC-606 Acetone/MeOH 33 Doc. Na N/D N/T N/D HPMC PhthalateDCM/MeOH 25 None N/D N/D N/T HPMC Phthalate DCM/MeOH 50 None N/D N/D N/THPMC Phthalate Acetone/MeOH 33 None Not harvested HPMC PhthalateAcetone/MeOH 33 None Not harvested HPMC Phthalate Acetone/MeOH 33 SLSN/D N/T N/D HPMC Phthalate Acetone/MeOH 33 Tween 80 Not harvested HPMCPhthalate Acetone/MeOH 33 Doc. Na N/D N/T N/D Eudragit L100-55 DCM/MeOH25 None N/D Present N/T Eudragit L100-55 DCM/MeOH 50 None N/D PresentN/T Eudragit L100-55 Acetone/MeOH 25 None N/D N/D N/T Eudragit L100-55Acetone/MeOH 50 None N/D N/D N/T Eudragit L100-55 Acetone/MeOH 33 SLSN/D N/T N/D Eudragit L100-55 Acetone/MeOH 33 Tween 80 N/D N/T N/DEudragit L100-55 Acetone/MeOH 33 Doc. Na N/D N/T N/D Eudragit E100DCM/MeOH 25 None N/D N/D N/T Eudragit E100 DCM/MeOH 50 None N/D N/D N/TEudragit E100 Acetone/MeOH 25 None N/D N/D N/T Eudragit E100Acetone/MeOH 50 None Present¹ N/T Present¹ Eudragit E100 Acetone/MeOH 33SLS N/D N/T N/D Eudragit E100 Acetone/MeOH 33 Tween 80 N/D N/T N/DEudragit E100 Acetone/MeOH 33 Doc. Na N/D N/T N/D Kleptose HP DCM/MeOH25 None N/D N/D N/T Kleptose HP DCM/MeOH 50 None N/D N/D N/T Kleptose HPAcetone/MeOH 25 None N/D N/D N/T Kleptose HP Acetone/MeOH 33 None N/DN/T N/D Kleptose HP Acetone/MeOH 50 None N/D N/D N/T Kleptose HPAcetone/MeOH 33 None N/D N/T N/D Kleptose HP Acetone/MeOH 33 None N/DN/T N/D Kleptose HP Acetone/MeOH 33 None N/D N/T N/D HPC Acetone/MeOH 25None N/D N/D N/T HPC Acetone/MeOH 50 None N/D N/D N/T CopovidoneAcetone/MeOH 25 None N/D N/D N/T Copovidone Acetone/MeOH 50 None PresentPresent N/T Kleptose/PVP K25 (70:5) Acetone/MeOH 25 None N/D N/T N/DKleptose/PVP K25 (45:5) Acetone/MeOH 50 None N/D N/T N/D Kleptose/PVPK25 (65:10) Acetone/MeOH 25 None N/D N/T N/D Kleptose/PVP K25 (40:10)Acetone/MeOH 50 None N/D N/T N/D Kleptose/HPMC-606 (70:5) Acetone/MeOH25 None N/D N/D N/T Kleptose/HPMC-606 (45:5) Acetone/MeOH 50 None N/DN/D N/T Kleptose/HPMC-606 (65:10) Acetone/MeOH 25 None N/D N/D N/TKleptose/HPMC-606 (40:10) Acetone/MeOH 50 None N/D N/D N/T Key: N/D =not detected N/T = not tested ¹Test performed in a separate study fromother Eudragit E100 entries

The results of the screening study demonstrate that preparation ofamorphous solid dispersions was possible for all of the polymersevaluated. However, solid dispersions produced using the low-meltingpoloxamers and polyethylene glycol were highly unstable, leading to theformation of crystalline drug within 1 month when stored at 30° C./60%relative humidity. No further evaluation of these polymers wasperformed. Solid dispersions produced with Eudragit E100 at 25% drugloading appeared to be amorphous and stable; however, crystallisationwas immediately apparent at 50% drug loading. Literature reportsindicate that dispersions produced with Eudragit E may exhibitsignificant crystallinity (e.g. see Qi et al. Int. J. Pharm.354:158-167, 2008); and, in a comparative study, may be less chemicallystable than solid dispersions produced using Povidone K25 (Dargel, E.,Mielck, J. B. Acta Pharm. Technol. 35(4):197-209. 1989). No furtherevaluation of Eudragit E100 was performed. Solid dispersions producedwith Eudragit L100-55 using a DCM/MeOH solvent system exhibitedcrystallisation after 1 week at 30° C./60% relative humidity, but thoseproduced using an acetone/MeOH solvent system were stable. We found thatsolid dispersions produced with copovidone at 50% drug loading exhibitedsome crystallisation after 1 week at 30° C./60% relative humidity, butthose produced at 25% drug loading were stable.

EXAMPLE 4. COMPOUND 1 FORMULATIONS 4.1 Immediate Release Tablet 4.1.1Composition

TABLE 7 Composition of an immediate release tablet % of core Ingredientmg/tablet weight Function Compound 1 100.00 25.00 Drug substance Lactose238.00 59.50 Filler Microcrystalline cellulose 40.00 10.00 FillerCroscarmellose Na 16.00 4.00 Disintegrant Sodium Lauryl Sulphate 2.000.50 Surfactant Magnesium stearate 4.00 1.00 Lubricant Core tabletweight 400.00

4.1.2 Method of Preparation

Standard immediate release tablets were manufactured using a directcompression process. Crystalline compound 1 and the lactose,microcrystalline cellulose, Croscarmellose Na and Sodium Lauryl Sulphatewere weighed into a glass vial to occupy approximately 75% of the volumeof the vial and then mixed together in a tumble mixer for 30 minutes.The blended material was sieved through a 40 mesh (425 μm) sieve, thentumble mixed for a further 15 minutes. The magnesium stearate was thenadded and the blend was shaken manually for about 20 seconds. Theresultant mixture was then dispensed into 400 mg portions and compressedinto tablet cores, using a hand press equipped with 10 mm tooling andwith a target compression force of 0.5 tonnes.

4.2 Microsuspension 4.2.1 Method of Preparation

Approximately 1 g of crystalline Compound 1 was weighed into a 10 mLvolumetric flask and 0.5% HPMC (hydroxypropyl methyl cellulose orHypromellose, USP substitution type 2910 having nominal apparentviscosity 4000 cP, such as DOW Methocel E4M or equivalent) solution wasadded to volume. The mixture was stirred overnight then quantitativelydiluted to 100 mL with 0.5% HPMC solution to give a 10 mg/mLmicrosuspension. The mean volume diameter of the Compound 1 wasdetermined to be 4.54 μm by laser diffraction using a Sympatec particlesize analyser (Sympatec GmbH).

4.3 Gelucire Capsule 4.3.1 Formulation

TABLE 8 Quantitative composition of Compound 1 50 mg capsules Amount perAmount Constituent capsule (mg) (% w/w) Function Standard Capsulecontents Compound 1 50.0 10.0 Active AstraZeneca Lauroyl 450.0 90.0Excipient, PhEur (NF^(c)) macrogolglyceride pharma- (Lauroyl ceuticalpolyoxylglyceride)^(a) aid Capsule Hypromellose Size 0 Each Dosage USP,Ph Eur capsule shell^(b) unit form presen- tation Titanium dioxide 1.84Each Opacifier unit Opacode black ink 0.0332 Each (S-1-7822/S-1-7823)unit ^(a)Supplied as Gelucire 44/14 grade. ^(b)Supplied as Capsugel VCap capsules

4.3.2 Method of Preparation

The lauroyl macrogolglyceride (lauroyl polyoxylglyceride) was melted atabout 50-70° C. then weighed into a stainless steel vessel. CrystallineCompound 1 was added and the contents mixed to achieve a homogeneoussuspension. Mixing was continued while the mixture was dispensed intocapsules to a fill weight of 500 mg per capsule using athermostatically-controlled automated capsule filling machine.

4.4 In Vitro Dissolution of Compound 1 Preparations 4.4.1 Test Method

Dissolution was performed according to the general procedure of theUnited States Pharmacopeia Apparatus I (Basket). An amount of materialcontaining approximately 100 mg of Compound 1 was weighed accuratelythen transferred to a dissolution vessel containing 500 mL of TRISbuffer (0.05M tris(hydroxymethyl)aminomethane solution adjusted to pH7.2 with hydrochloric acid) maintained at 37° C. and stirred at 100 rpm.After 15, 30, 45 and 60 minutes, 10 mL samples were withdrawn andfiltered through 0.2 μm PVDF filters. Compound 1 concentration in thefiltrate was determined by ultraviolet spectroscopy at a wavelength of278 nm.

4.4.2 Results

TABLE 9 In vitro dissolution of Compound 1 preparations Dissolution (%Release) 15 30 45 60 75 90 105 120 Sample min min min min min min minmin Drug only 15 28 43 51 58 62 68 71 Tablet 72 81 85 87 89 90 91 92Micro- 70 75 77 78 79 79 80 80 suspension Gelucire 37 92 97 99 99 100100 100 capsule (10% drug loading)

See FIG. 2. 4.5 Nanosuspension 4.5.1 Method of Preparation

Compound 1 was mixed with a few drops of vehicle (0.5% HPMC/0.1%Tween80) in a glass vial and “vortex” mixed for 1 minute, to wet anddisperse the compound and to form a free flowing slurry. A furthervolume of vehicle was added to the slurry to produce a drugconcentration of 50 mg/ml and the resulting slurry was then “vortex”mixed for approximately 1 minute to mix. The slurry at 50 mg/ml drugconcentration was transferred to a zirconia milling pot. Zirconiamilling beads (0.6-0.8 mm diameter) were added to the pot until thelevel of beads and slurry was equal. The pot was then sealed with aTeflon ring and lid (zirconia) and placed on a Fritsch P7 planetarymill. A second pot (as counter weight) was then placed on the mill. Thepots were rotated on the mill at 800 rpm for 4×30 minutes runs (with 10minutes between each run). The pots were then allowed to cool for afurther 15 minutes and a sample of the resulting bead milled suspensiontaken for analysis. The nanosuspension was then separated from themilling beads, and diluted to a concentration of 10 mg/ml, ready fordosing. Nanosuspension particle size was measured using Fibre OpticQuasi Elastic Light Scattering (FOQUELS) from BrookhavenInstruments—laser wavelength of 635 nm. A mean effective diameter of692+/−8 nm was measured. X-ray diffraction confirmed that the drug wasessentially crystalline.

4.6 Solid Dispersion 4.6.1 Preparation by Solvent Evaporation Process

Solid dispersions having a 1:3 ratio by weight of Compound 1:polymerwere prepared as follows:

0.75 g of Compound 1, prepared according to Example 9 [compound 168] inWO 2004/080976, and 2.25 g of polymer were weighed directly into a 250ml round bottom flask and dissolved in 75 ml of methanol:dichloromethane(1:1). The solvent was removed on a rotary evaporator. The formulationwas placed in a vacuum oven and dried under high vacuum at 40° C.overnight.

The formulation was retrieved from the flask and dry milled if necessaryusing a pestle and mortar. The formulation was then stored in a vacuumdesiccator until needed.

In order to produce formulations having ratios other than 1:3, weightsand volumes in the process were adjusted pro-rata to those describedabove.

4.6.2 Preparation by Melt Extrusion Process

Compound 1 was blended with polymer and glidant in the proportionsdefined in the manufacturing formula. The blend was extruded in atwin-screw extruder. During extrusion, a vacuum was applied to theextruder barrel to degas the melt. The extrudate was calendered bypassing through two contra-rotating calender rollers, and then cooledprior to milling.

4.6.3 Stability Study 4.6.3.1 Protocol

Solid dispersions were prepared using the solvent evaporation processdescribed previously (see 4.6.1), and amorphous Compound 1 was preparedaccording to Example 9 [compound 168] in WO 2004/080976. Samples werestored in closed HDPE bottles with polyethylene liners, with desiccant,for a period of 3 months under refrigeration (2-8° C.), long-termconditions (25° C./60% relative humidity) and accelerated conditions(40° C./75% relative humidity). In addition, samples were stored for aperiod of 1 month in an open petri dish at 40° C./75% relative humidity.Samples were tested prior to set-down, after 1 month and, for thesamples in closed containers under long-term and accelerated conditionsonly, after 3 months.

4.6.3.2 Methodology Dissolution

Dissolution was carried out in accordance with the general procedure ofthe United States Pharmacopeia using Apparatus II (paddle method). Anamount of the solid dispersion containing about 100 mg of Compound 1 wasweighed accurately then placed in 500 mL pH6.5 phosphate buffer at atemperature of 37° C. and a stirring speed of 75 rpm. After 5, 10, 20and 45 minutes a 2 mL sample was removed and the Compound 1 contentdetermined by HPLC.

TABLE 10 Chromatographic conditions for in vitro dissolution testApparatus Liquid chromatograph with UV detector Column Waters SunfireC18, 4.6 mm × 50 mm (3.5 μm or equivalent) Eluents Eluent A: 0.1% TFA inwater Eluent B: 0.1% TFA in acetonitrile Time (min) % A % B Gradientprogram 0 65 35 0.8 65 35 0.81 5 95 1.8 5 95 1.81 65 35 3.5 65 35 Flowrate 1 mL/min approx. Temperature 35° C. Wavelength 276 nm Injectionvolume 10 μL Run time 3.5 min. Compound 1 1 min approx. retention time

Determination of Crystallinity by Differential Scanning Calorimetry

The sample was heated in a differential scanning calorimeter (TAInstruments Q1000) using a programme designed to drive off any waterand/or solvents present, before cooling the sample and heating at aconstant rate over a temperature range encompassing the meltingtransition of any crystalline material which may be present (Compound 1Tm=210° C.) (see FIG. 3).

TABLE 11 Parameters for differential scanning calorimetry Generalparameters Sample weight (mg) 2-10 Pan type Aluminium, piercedAtmosphere Nitrogen, 20-30 mL/min Temperature programme Equilibration(30 minutes) 30° C. Cool to 0° C. Heat at 5° C./min 120° C. Cool 0° C.Heat at 5° C./min 235° C. Cool

4.6.3.3 Results

TABLE 12 Results for the stability study of Compound 1 polymericdispersions 2-8° C. 25° C./60% RH 40° C./75% RH Closed Closed ClosedOpen Initial 1 month 1 month 3 months 1 month 3 months 1 monthFormulation Diss DSC Diss DSC Diss DSC Diss DSC Diss DSC Diss DSC DissDSC Kleptose 1:3 90 N/D 88 N/D 91 N/D 92 N/D 87 N/D 84 N/D NT N/D PVP1:3 92 N/D 91 N/D 91 N/D 94 N/D 90 N/D 66 X NT X Amorphous NT N/D NT XNT X NT X NT X NT X NT X Compound 1 Kleptose 1:2 81 NT 82 N/D 82 N/D XN/D 76 N/D 66 N/D 81 N/D PVP 1:2 81 N/D 81 N/D 77 N/D 86 N/D 85 N/D 55N/D NT X HPMCP 1:3 99 N/D 91 N/D 90 N/D 87 N/D 87 N/D 83 N/D 91 N/DHPMCP 1:2 97 N/D 98 N/D 97 N/D 92 N/D 91 N/D 89 N/D 92 N/D Key: N/D =not detected N/T = not tested Diss = Dissolution (cumulative release) at45 minutes, % DSC = Crystallinity as determined by differential scanningcalorimetry

The results of the stability study demonstrate that solid dispersionsproduced using the relatively hygroscopic polymer povidone tended tocrystallise when stored at 40° C./75% relative humidity, leading to areduction in dissolution rate. Solid dispersions produced using2-hydroxypropyl-β-cyclodextrin and hypromellose phthalate were stableunder all tested conditions.

4.7. Copovidone Solid Dispersion (Uncoated Tablet Formulation) 4.7.1Formulation

TABLE 13 Composition of Compound 1/copovidone solid dispersion uncoatedtablet Quantity Quantity Components (mg) (%) Function Standard Compound1 200.00 25.00 Active AstraZeneca pharma- ceutical ingredient Copovidone460.00 57.50 Polymeric NF and Ph Eur carrier Colloidal 14.64 1.83Glidant NF and Ph Eur silicon dioxide Mannitol 117.36 14.67 Soluble NFand Ph Eur filler Sodium stearyl 8.00 1.00 Lubricant NF and Ph Eurfumarate Core tablet 800.00 weight

4.7.2 Method of Preparation

A solid dispersion of Compound 1 and copovidone was prepared using themelt extrusion process described in 4.6.2. The milled extrudate wasmixed with the external excipients and compressed into tablet form usinga single punch hand press to achieve hardness in the range 80-100 N.

4.7.3 Stability Study—Uncoated Tablets 4.7.3.1 Protocol

Uncoated tablets prepared as described in 4.7.2 were stored in closedHDPE bottles with polyethylene liners, with desiccant, for a period of 4months under long-term conditions (25° C./60% relative humidity) andaccelerated conditions (40° C./75% relative humidity). Samples weretested prior to set-down, then after 1, 3 and 4 months.

4.7.3.2 In Vitro Evaluation

Crystallinity was determined by DSC as described in 4.6.3.2.

Dissolution Test

The dissolution method was adapted from that previously described forsolid dispersion formulations (see 4.6.3.2). Dissolution was carried outin accordance with the general procedure of the United StatesPharmacopeia using Apparatus II (paddle method). Individual dosage unitswere placed in 1000 mL of pH6.5 phosphate buffer at a temperature of 37°C. and a stirring speed of 75 rpm. After 15, 30, 60, 90, 120 and 180minutes a 1 mL sample was removed and the Compound 1 content determinedby HPLC:

TABLE 14 Chromatographic conditions for in vitro dissolution test forCompound 1/copovidone solid dispersion tablet Chromatographic conditionsApparatus Liquid chromatograph with UV detector Column Waters SunfireC18, 4.6 mm × 50 mm (3.5 μm or equivalent) Eluents Eluent A: 0.1% TFA inwater Eluent B: 0.1% TFA in acetonitrile Time (min) % A % B Gradientprogram 0 75 25 3.0 55 45 3.5 0 100 4.0 0 100 7.0 75 25 Flow rate 1mL/min approx. Temperature 40° C. Wavelength 276 nm Injection volume 10μL Run time 7 min. Compound 1 2.9 min approx. retention time

Compound 1 Assay and Impurities by HPLC

The Compound 1 and total impurities contents were determined using HighPerformance Liquid Chromatography (HPLC). A sample solution was preparedcontaining approximately 0.4 mg/mL Compound 1, using 50:50 v/vacetonitrile/water as diluent. The sample solution was filtered using a0.2 μm PVDF filter prior to analysis.

10 μL sample was injected into a mobile phase comprising 0.05%trifluoroacetic acid (TFA) in water (Eluent A)/0.05% TFA in acetonitrile(Eluent B), as defined by the gradient program in Table 15 below.

TABLE 15 Gradient programme - Compound 1 assay and impurities Gradientprogramme Time mins) % A % B 0 90 10 20 60 40 28 5 95 30 5 95 30.1 90 1036 90 10

The mobile phase starts as defined at time zero, then the composition ismodified by adjusting the proportion of eluents A and B gradually andlinearly to the composition at each successive time-point.

Separation of impurities was performed using a column 15 cm long×4.6 mminternal diameter packed with Waters Sunfire C18 stationary phase having3.5 μm particle size. The mobile phase flow rate was 1.0 mL/minute,temperature was controlled at 30° C., and impurity concentration wasdetermined by comparison of absorbance at 276 nm, measured using avariable wavelength uv detector, with that of an external Compound 1reference standard.

Water Content by Coulometric Karl Fischer Titration

Water content was determined by coulometric Karl Fischer titration usinga Metrohm 684 Coulometer. Samples were ball milled prior to analysis andmeasurements were performed using a sample size of 200 mg.

4.7.3.3 Results

TABLE 16 Results of the stability study for Compound 1/copovidone soliddispersion tablet (200 mg, uncoated)) 25° C./60% Relative Humidity 40°C./75% Relative Humidity Initial 1 month 3 months 4 months 1 month 3months 4 months Crystallinity by DSC N/D N/D N/D N/D N/D N/D N/DDissolution (Time-point) X¹ S² X¹ S² X¹ S² X¹ S² X¹ S² X¹ S² X¹ S² (15min) 14 3 15 2 19 7 20 5 17 2 14 1 17 2 (30 min) 32 5 33 3 41 15 45 1038 2 33 3 37 4 (60 min) 60 8 62 4 68 13 81 15 70 2 62 7 68 5 (90 min) 775 82 8 85 6 96 7 88 3 80 7 85 2 (120 min)  84 2 89 6 92 3 100 4 93 5 885 91 2 (180 min)  87 1 91 4 93 1 NT 95 4 91 4 94 1 Water content (% w/w)1.3 1.3 1.6 1.3 1.4 1.7 1.8 Assay (%) 99.6 98.6 101.1 98.1 100.4 100.5100.1 Impurities (%) 0.44 0.44 0.44 0.43 0.44 0.44 0.44 ¹X is the mean %release (n = 3) ²S is the standard deviation (n = 3)

4.8. Copovidone Solid Dispersion (Film-Coated Tablet Formulation) 4.8.1Formulation

TABLE 17 Composition of Compound 1/copovidone solid dispersion tabletComponents 25 mg 100 mg tablet tablet Quantity (mg Quantity (% Tabletcore per tablet) core weight) Function Compound 1 25.00 100.00 25.00Active pharmaceutical ingredient Copovidone 57.50 230.00 57.50 Polymericcarrier Colloidal 1.83 7.33 1.83 Glidant silicon dioxide Mannitol 14.6758.67 14.67 Soluble filler Sodium 1.00 4.00 1.00 Lubricant stearylfumarate Core tablet 100.00 400.00 weight Quantity (mg Quantity (%Tablet Coating per tablet) coating weight) Function Hypromellose 2.198.75 62.5 Film former (HPMC 2910) Titanium 0.88 3.51 25.05 Opacifierdioxide (E171) Macrogol/ 0.22 0.88 6.25 Plasticiser PEG 400 Components25 mg 100 mg tablet tablet Quantity (mg Quanity (% Tablet core pertablet) core weight) Function Iron oxide 0.16 0.64 4.55 Colouring agentyellow (E172) Iron oxide 0.06 0.23 1.65 Colouring agent black (E172) %of core weight Nominal 3.50 14.00 3.50 Coating Weight

4.8.2 Method of Preparation

Compound 1 was blended with polymer and glidant in the proportionsdefined in the manufacturing formula. The blend was extruded in atwin-screw extruder. During extrusion, a vacuum was applied to theextruder barrel to degas the melt. The extrudate was calendered bypassing through two contra-rotating calender rollers, and then cooledprior to milling. The extrudate was milled and subsequently mixed withthe external excipients. The powder blend was compressed into tabletcores using a Rotary Press (Korsch XL 100 with 10 punch stations) toachieve a sufficient hardness (minimum 25 N).

The tablet cores were coated using a Driacoater Driam 600 coater withOpadry™ Green (Colorcon 03B21726, 130 g/Kg aqueous solution). The totalcoating solution applied is equivalent to 35 g of Opadry™ per Kg oftablet cores.

4.8.3 Stability Study—Film-Coated Tablets 4.8.3.1 Protocol

Film-coated tablets prepared as described in 4.8.2 were stored in closedHDPE bottles with polyethylene liners, with desiccant, for a period of 4months under long-term conditions (25° C./60% relative humidity) andaccelerated conditions (40° C./75% relative humidity). Samples weretested prior to set-down, then after 1 month 3 and 4 months.

4.8.3.2 In Vitro Evaluation

Water content, assay and impurities were determined using the methodsdescribed in Section 4.7.3.2.

Determination of Crystallinity by Hot-Stage Microscopy

Ground tablets were examined by optical microscopy undercross-polarising conditions whilst being heated steadily across themelting point range of the excipients and Compound 1 to detect thepresence of drug crystals. Any particles seen to be birefringent between180° C. and 190° C. which subsequently melted at approximately 210° C.were classified as Compound 1. See FIG. 4 for an example of a drugcrystal as seen under the microscope.

Dissolution Test

The dissolution method was adapted from that previously described foruncoated tablet formulations (see 4.7.3.2). Dissolution was carried outin accordance with the general procedure of the United StatesPharmacopeia using Apparatus I (basket method). Individual dosage unitswere placed in 900 mL 0.3% SDS at a temperature of 37° C. and a stirringspeed of 100 rpm. After 15, 30, 45, 60 and 90 minutes a sample wasremoved and the Compound 1 content determined by HPLC:

TABLE 18 Chromatographic conditions for in vitro dissolution test forCompound 1/copovidone solid dispersion tablet Chromatographic conditionsApparatus Liquid chromatograph with UV detector Column Waters SymmetryC18, 4.6 mm × 75 mm × 3.5 μm Eluents Eluent A: 0.1% TFA in water EluentB: 0.1% TFA in acetonitrile Time (min) % A % B Gradient program 0 75 253.0 55 45 3.5 0 100 7.0 75 25 Flow rate 1 mL/min approx. Temperature 40°C. Wavelength 276 nm Injection volume 10 μL Run time 7 min Compound 12.9 min approx. retention time

4.8.3.3 Results

TABLE 19 Results of the stability study for Compound 1/copovidonefilm-coated solid dispersion tablet (25 mg) 25° C./60% Relative Humidity40° C./75% Relative Humidity Initial 4 weeks 13 weeks 26 weeks 4 weeks13 weeks 26 weeks Crystallinity: D (+) N/D D (++) D (+++) N/D D (++) D(+++) Hot-Stage Microscopy N/D N/T N/D N/D N/T N/D N/D Wide-Angle X-RayScattering Dissolution (Time-point) X¹ S² X¹ S² X¹ S² X¹ S² X¹ S² X¹ S²X¹ S² (15 min) 41 3.6 38 3.2 41 3.8 41 2.9 39 2.8 41 2.1 39 3.5 (30 min)77 5.2 78 6.2 78 4.8 81 4.5 77 3.7 78 2.1 78 5.4 (45 min) 98 3.9 99 3.599 3.4 102 2.3 98 3.9 98 1.4 101 2.4 (60 min) 104 1.4 104 1.9 104 1.0105 1.3 103 4.8 101 0.5 106 1.3 (90 min) 104 1.1 104 1.4 104 1.0 105 1.5103 4.5 101 0.4 106 1.0 Water content (% w/w) 2.3 2.1 2.2 2.0 1.9 2.12.2 Assay (%) 104.0 104.3 103.5 102.5 102.0 104.1 106.0 Impurities (%)0.52 0.51 0.50 0.50 0.50 0.50 0.53 Key: N/D = not detected D = detected;(+) 1-5 birefringent spots (++) 5-30 birefringent spots (+++) more than30 birefringent spots N/T = not tested ¹X is the mean % release (n = 3)²S is the standard deviation (n = 3)

TABLE 20 Results of the stability study for Compound 1/copovidonefilm-coated solid dispersion tablet (100 mg) 25° C./60% RelativeHumidity 40° C./75% Relative Humidity Initial 4 weeks 13 weeks 26 weeks4 weeks 13 weeks 26 weeks Crystallinity: D (+) N/D D (+++) D (+++) D (+)D (+) D (++) Hot-Stage Microscopy N/D N/T N/D N/D N/T N/D N/D Wide-AngleX-Ray Scattering Dissolution (Time-point) X¹ S² X¹ S² X¹ S² X¹ S² X¹ S²X¹ S² X¹ S² (15 min) 24 0.5 24 1.0 25 1.9 26 1.1 25 1.8 25 1.2 24 1.2(30 min) 55 1.0 54 1.3 56 2.3 60 1.6 57 2.8 56 2.1 56 1.9 (45 min) 801.6 80 1.6 81 1.9 87 1.5 83 3.1 81 2.1 83 2.1 (60 min) 97 1.0 97 1.1 981.7 102 0.5 99 2.1 97 2.1 99 1.2 (90 min) 101 0.8 101 0.5 102 0.8 1040.8 102 1.0 101 0.8 102 0.5 Water content (% w/w) 2.0 1.7 2.5 1.6 1.82.2 1.5 Assay (%) 102.5 100.5 102.8 102.2 103.6 100.8 102.1 Impurities(%) 0.50 0.49 0.50 0.50 0.51 0.49 0.50 Key: N/D = not detected D =detected; (+) 1-5 birefringent spots (++) 5-30 birefringent spots (+++)more than 30 birefringent spots N/T = not tested ¹X is the mean %release (n = 3) ²S is the standard deviation (n = 3)

EXAMPLE 5 NANOMETER-SCALE CHARACTERISATION STUDIES

5.1 Solid state Nuclear Magnetic Resonance Study

Solid dispersions of Compound 1 and copovidone, prepared with drugloadings of 10, 25, 35 and 40% using the melt extrusion processdescribed in 4.6.2, were evaluated by solid state nuclear magneticresonance spectroscopy (SSNMR) using methodology disclosed in Asano, A;Takegoshi, K.; Hikichi, K. Polymer (1994), 35(26), 5630-6. ¹³Ccross-polarisation magic angle spinning SSNMR spectra were recorded at100 MHz with a spin rate of 9 kHz using a Bruker Avance 400WB with a 4mm HFX MAS probe. For each sample, with different drug loading, a seriesof spectra were acquired with different contact times ranging from 500μs to 10 ms. Peak areas from different spectral regions were measured.These areas were selected to contain peaks corresponding to Compound 1or copovidone. With increasing contact time peak area increases to amaximum value and then decays due to a process known as proton spindiffusion. This decay is characterised by a constant T_(1ρ), whichrepresents proton spin-lattice relaxation in the rotating frame ofreference. For a phase-separated system, on a length scale longer thanthe spin-diffusion length scale, the relaxation rates for this decayprocess are identical to those observed for the individual components.For a mixed system, a single value of T_(1ρ) is observed as a weightedaverage of the individual components.

For the samples with Compound 1 loading between 10 & 40% eachmagnetization decay could be fitted to a single exponential functionwith very similar T_(1ρ) values observed. This suggests a similarrelaxation pathway for the drug and polymer and infers a single phase.

TABLE 21 Results of the Solid State NMR study T_(1ρ)/ms Compound 1 Peaksdue to Compound 1 Peaks due to co-povidone loading (119.5-140.0 ppm)(167.0-183.0 ppm) 40% 9.9 9.7 35% 10.2 9.4 25% 13.2 8.6 10% 15.5 9.4

5.2 Pair-Wise Distribution Function Study

Solid dispersions of Compound 1 and copovidone, prepared with drugloadings of 10, 25, 35 and 40% using the melt extrusion processdescribed in 4.6.2, were evaluated using X-ray powder diffraction andPair-wise Distribution Functions (PDFs) were derived for each sample.

5.2.1 Data Collection

X-ray powder diffraction data were collected on the Bruker D8diffractometer, which has a Copper source generating X-rays with awavelength of 1.5418 Å (Gobel mirrors used to provide parallel beamoptics remove the kβ leaving a beam with an average wavelength of kα1and kα2) using a voltage of 40 kV and a filament emission of 40 mA.Samples were measured in reflection mode and the diffraction patterncollected using a scanning position-sensitive detector.

A diffractogram of the zero background wafer was obtained, under vacuum.50 mg (+/−5 mg) of each sample was weighed out and dispersed onto a zerobackground holder, ensuring near complete coverage. The sample was addedto the TTK chamber, which was then placed under vacuum to a pressure of<5×10−2 mbar. XRPD data were collected over approximately 20-30 minutes:data acquisition parameters of 4-80°2θ in steps of 0.007091° countingfor 0.2 s/step were used for each sample.

A peak in the patterns at 6.6°2θ is caused by the sample holder and wasremoved in each case through subtraction of a blank run (i.e. an emptysample holder) which is measured on the day of the experiment.

5.2.2 Computational Methods—Pair-Wise Distribution Function

PDFs were calculated for each sample (S. J. L. Billinge and M. G.Kanatzidis, Chem. Commun., 2004, 749-760; S. Bates et. al.,Pharmaceutical Research, 2006, 23(10) 2333-2349; S. Bates et. al., J.Pharmaceutical Sciences, 2007, 96(5), 1418-1433) The measured X-raydiffraction pattern (known as the scattering function) was transformedto a normalized scattering function S(Q) by carrying out a number ofcorrections to the data related to both the sample and experimentalset-up. PDFs are then generated from the sine Fourier transformation ofs(Q), equation 1.

$\begin{matrix}{{G(r)} = {\frac{2}{\pi}{\int_{0}^{\infty}{{Q\left\lbrack {{S(Q)} - 1} \right\rbrack}{\sin ({Qr})}\ {dQ}}}}} & {{Equation}\mspace{14mu} 1}\end{matrix}$

Q is the magnitude of the scattering vector and is derived from Q=4πsin(q)/λ

The PDF is a plot of G(r) against interatomic distance and shows theprobability of finding an atom at given distance ‘r’ from another atom.X-ray amorphous materials which are nanocrystalline possess long rangeordering and thus the probability of finding an atom out at longdistances is relatively high. In contrast, truly amorphous materials donot possess any long range ordering and the probability of finding anatom out at long distances is relatively low.

PDFs were generated from each of X-ray diffraction pattern measuredusing the software PDFgetX2 (X. Qui et. al., J. Appl. Cryst. 2004, 37,678)

5.2.3 Results

As shown in FIG. 5. there is little evidence of ordering beyond 15 Å forsolid dispersions of Compound 1 and copovidone for any of the drugloadings investigated. This confirms that these solid dispersions areamorphous and do not exhibit significant long-range order.

5.2.4 Linear Combination of PDFs 5.2.4.1 Method

PDFs of the separate components of the formulation, amorphous Compound 1and copovidone, were generated. These PDFs were then combined in thecorrect ratios (70% copovidone and 30% amorphous Compound 1) to give asimulated PDF trace for a physical mixture of the two. The tracesobtained in 5.2.2. were compared to this simulated trace.

5.2.4.2 Results

As shown in FIG. 6, a physical mixture of amorphous Compound 1 andcopovidone would exhibit a characteristic pattern between 1 and 5 Å,comprising dual minima for G(r) at approximately 2 Å and approximately 3Å; solid dispersions of Compound 1 and copovidone exhibit a singleaccentuated minimum at approximately 3 Å. These data indicate that soliddispersions of Compound 1 and copovidone are true molecular dispersions.

5.3 Nano-Thermal Characterisation Study

Solid dispersions of Compound 1 and copovidone, prepared with drugloadings of 10, 30 and 40% using the melt extrusion process described in4.6.2, were evaluated using Atomic Force Microscopy (Gan, Y. SurfaceScience Reports (2009), 64(3), 99-121; Fulghum, J. E.; McGuire, G. E.;Musselman, I. H.; Nemanich, R. J.; White, J. M.; Chopra, D. R.;Chourasia, A. R. Analytical Chemistry (1989), 61(12), 243R-69R) andusing localised thermal analysis (Harding, L.; King, W. P.; Dai, X.;Craig, D. Q. M.; Reading, M. Pharmaceutical Research (2007), 24(11),2048-2054.)

5.3.1 Methods

The work was carried out on a TA Instruments 2990 Micro-Thermal Analyzerbased on a Veeco Explorer atomic force microscope. Preliminary imagingof the samples was carried out in Tapping Mode (TM-AFM) using Veeco1660-00 high resonance frequency (HRF) silicon probes. Micro-thermalanalysis (micro-TA) was carried out using Wollaston wire thermal probes.Nano-thermal analysis (nano-TA) was carried out using Anasys InstrumentsAN2-300 ThermaLever™ doped silicon probes controlled by an AnasysInstruments NanoTA1 AFM accessory. The Wollaston probe wastemperature-calibrated using poly(ethylene) terephthalate (PET) film(melting temperature=240° C.) and room temperature. A 3-pointtemperature calibration was carried out for the ThermaLever probe usingpolycaprolactone (PCL, Tm=55° C.), HDPE (Tm=115° C.) and PET meltingtemperature standards. The calibration of each probe was checked beforeand after a sample was analysed. Unless stated otherwise, the heatingrate used in all the localised thermal analysis runs was 20° C./s.

All the samples were analysed in the as-received state—i.e. theunmodified surface of the moulded pellets.

5.3.2 Results

The samples at various drug loadings all exhibited surface features to avariable degree, but none showed any evidence of phase separation withinthe matrix, as illustrated in FIG. 7 (10% drug loading), FIG. 8 (30%drug loading) and FIG. 9 (40% drug loading).

5.4 Crystallisation Study

The effect of water on the crystallinity of Compound 1 was investigatedfor the milled extrudate prepared using the melt extrusion processdescribed in 4.6.2 and for the tablet composition shown in Table 13 andprepared as described in 4.7.2. The study was carried out using aqueousslurries both in the absence and presence of a proprietary coatingcomposition, Opadry™ Green (Colorcon 03B21726, 130 g/Kg aqueoussolution). Tablets were ground prior to the slurry experimentscommencing.

5.4.1 Experimental Conditions

The following materials were weighed into 25 mL vials:

TABLE 22 Preparation of slurries for crystallisation study Weight (mg)Exp Ground tablet Milled extrudate Opadry 1 83.0 199.2 2 — 67.7 208.2 391.0 — — 4 — 68.0 —

20 mL of water heated to 50° C. was added to each vial. The resultingslurries remained stirring at 50° C. for 48 hours.

Analysis of the resultant slurry material by XRPD identified Form H asthe primary crystal form of Compound 1. Compound 1 Form H has an X-raydiffraction pattern (λ=1.5418 Å) containing specific peaks at:

TABLE 23 XRPD data for Compound 1 Form H Peak 2θ° (±0.1°) 1 6.5 2 6.9 38.4 4 12.8

Compound 1 Form H may also have the following additional peaks an X-raydiffraction pattern (λ=1.5418 Å):

TABLE 24 Additional XRPD data for Compound 1 Form H Peak 2θ° (±0.1°) 515.1 6 16.5 7 16.8 8 19.9 9 20.3

Compound 1 Form H may also be characterised by any combination of threeor more peaks selected from the first list of 4 peaks above.

A representative powder XRPD pattern of compound 1 Form H is shown inFIG. 10.

Compound 1 Form H gives a weight loss via TGA that is consistent with amonohydrate with some additional physisorbed water. In the example giventhe total amount of water present is 4.7% by weight; the theoreticalweight loss for a monohydrate of compound 1 is 4.0% w/w.

Compound 1 Form H may also be characterised using DSC. Compound 1 Form Hwhen heated from 0° C. to 300° C. at 10° C. per minute displays a broaddehydration endotherm up to 115° C., followed by phase transitionsbetween 125-175° C. A sharp endotherm is observed with an onset at208.0° C.±1° C., this being consistent with Form A. A representative DSCtrace for compound 1 as Form H is shown in FIG. 11.

In the absence of Opadry™ the resulting material gave strong XRPDreflections consistent with Form H, whereas in the presence of Opadry™the intensity of the Form H XRPD diffraction pattern was considerablyreduced. This is not the result of interference, as the XRPD diffractionpattern of Opadry™ shown in FIG. 12 indicates there are no significantpeaks present below 25°2θ. Therefore, the very low intensity of thereflections observed indicates the presence of only small quantities ofForm H. This may suggest that Opadry™ may exert a stabilising effectupon amorphous solid dispersions of Compound 1. This grade of Opadry™was selected for use in the preparation of the film-coated tabletformulation described in 4.8.

5.5 Two-Dimensional Correlation Spectroscopy Study 5.5.1 Introduction

Two-dimensional correlation spectroscopy (2D-COS) is a method in whichan external perturbation is applied to a system and monitored by somespectrometric device. Spectral intensity is plotted as a function ofspectral variables (e.g. wavelength, frequency or wavenumber). Twoorthogonal axes of spectral variables define the 2D spectral plane, andthe spectral intensity may be plotted along a third axis (Noda, I.,Dowrey, A. E., Marcott, C., Story, G. M., Ozaki, Y. Appl. Spectrosc. 54(7) 2000 pp 236A-248A; Noda, I. Appl. Spectosc. 44 (4) 1990 pp 550-561).

In a synchronous 2D correlation spectrum, intensity is representative ofthe simultaneous or coincidental changes of spectral intensityvariations measured across the range of perturbation. A synchronousspectrum is symmetrical with regard to the diagonal corresponding toequal values for the chosen spectral variable; correlation peaks appearat both diagonal and off-diagonal positions. The diagonal peaks,referred to as autopeaks, represent the intensity variation for specificvalues of the chosen spectral variable across the range of perturbation,whereas the off-diagonal peaks, referred to as cross peaks, representsimultaneous or coincidental changes of spectral intensities observed attwo different values of the chosen spectral variable. Such synchronisedchanges may indicate a coupling or interaction.

In contrast, in the asynchronous spectrum, intensity representssequential or successive changes. The asynchronous spectrum isanti-symmetrical with respect to the diagonal and has no autopeaks,consisting exclusively of cross peaks which develop only if the twospectral features change out of phase. This feature may be used todifferentiate overlapped bands arising from spectral signals ofdifferent origins, such as different components acting independently ina complex mixture.

For both synchronous and asynchronous correlation spectra, sensitivitymay be improved, at the expense of an increase in noise, by subtractionof the average spectrum from each individual spectrum in theperturbation data set

Thus 2D-COS may be used to establish the nature and extent of anycorrelations in the spectral variations which arise in response to theperturbation and which may be indicative of intra- or inter-molecularinteractions within the sample matrix. In the context of apharmaceutical solid dispersion, a high level of interaction between thedrug and the matrix polymer will tend to promote the formation of astable and homogeneous dispersion whereas the absence of suchinteraction, or the existence of competitive intramolecular coupling,will have a contrary effect.

5.5.2 Method

The effect of a change in concentration of Compound 1 and variouspolymers in solid dispersions prepared by the solvent evaporationprocess described in 4.6.1 was studied using infrared spectroscopy. Thespectra were collected on a Thermo Nicolet Magna 550 series IIspectrometer. 2D-COS spectra were collected for solid dispersioncompositions of Compound 1 and matrix polymers as shown in Table 25.

TABLE 25 List of polymers with percent of mixtures Matrix polymerHypromellose Composition acetate succinate Copovidone HypromelloseHypromellose Povidone API % Polymer % (Aqoat MG) (Kollidon VA64)phthalate (HP55S) (Pharmacoat 606) (Kollidon 25) 10 90 T T T T T 20 80 TT T T T 23 77 T T T T T 25 75 T T T T T 26 74 T T T T T 28 72 T T N/T TT 30 70 T T N/T T T T = tested N/T = not tested

Each spectrum was normalised to the most intense band using proprietarysoftware (Omnic 8.0). The spectra were then converted into a commaseparated value (CSV) file, transferred to MS Excel™ and formatted forMatlab® (The MathWorks™) wherein 2D synchronous and asynchronous spectrawere generated.

5.5.3 Results Hypromellose Acetate Succinate (Aqoat MG)

In the spectrum of Compound 1, the most intense band is located at 1630cm⁻¹ (FIG. 13). In the Aqoat MG spectrum the most intense band islocated at 1050 cm⁻¹ (FIG. 14). In the synchronous spectrum (FIG. 15)cross peaks are evident at 1050 cm⁻¹, 1650 cm⁻¹ and 1050 cm⁻¹, 2700cm⁻¹; however the asynchronous spectrum (FIG. 16) indicates that theseinteractions are intramolecular (polymer/polymer) in nature.

Hypromellose Phthalate (HP55S)

The Infrared spectrum for HP55S exhibits a strong spectral feature atjust above 1000 cm⁻¹, as shown in FIG. 14. The synchronous (FIG. 17) andasynchronous (FIGS. 18 and 19) correlation spectra indicate weak mixedintra- and inter-molecular interactions in the range 1600 to 1800 cm⁻¹.

Hypromellose (Pharmacoat 606)

As for HP55S, the infrared spectrum for Pharmacoat exhibits a strongspectral feature at just above 1000 cm−1, (FIG. 14). The synchronous(FIG. 20) and asynchronous (FIGS. 21 and 22) correlation spectraindicate weak mixed intra- and inter-molecular interactions in the range1600 to 1800 cm⁻¹. The intensity of intermolecular (drug-polymer)interaction for Pharmacoat is somewhat greater than for HP55S.

Povidone (Kollidon 25)

The primary band in the infrared spectrum of povidone (FIG. 14) is at1600 cm⁻¹ and overlaps with the primary band in the infrared spectrum ofCompound 1 (FIG. 13). The synchronous (FIGS. 23 and 24) and asynchronous(FIG. 25) correlation spectra indicate hydrogen bonding interactions.

Copovidone (Kollidon VA64)

Copovidone has many of the same infrared (FIG. 2) and 2D spectralfeatures (FIGS. 26-29) as Povidone but also exhibits additional factorssuggesting a greater strength of hydrogen bonding.

5.5.4 Conclusions

The degree of intermolecular interaction observed in solid dispersionsof Compound 1 is highly dependent upon the nature of the matrix polymer.The overall ranking of the intermolecular interactions is shown in Table26.

TABLE 26 Molecular Interaction Ranking Polymer Interaction Strength RankAqoat MG Dipole-dipole Very Weak 5 HP55S Dipole-dipole Weak 4 PharmacoatDipole-dipole Medium to Weak 3 Povidone Hydrogen bonding Strong 2Copovidone Hydrogen bonding Very Strong 1

These results suggest that solid dispersions of Compound 1 andcopovidone may be particularly stable and homogeneous.

EXAMPLE 6. COMPARATIVE BIOAVAILABILITY STUDY 6.1 Protocol

One hundred milligrams of the drug in several different presentationswere orally administered to fasted beagle dogs (n=6). The formulationsdosed were the IR tablet (see 4.1), microsuspension (see 4.2) andnanosuspension (see 4.5) formulations, capsules containing various drugloadings in Gelucire® 44/14 (see 4.3), capsules containing soliddispersions produced by solvent evaporation (see 4.6.1), and meltextrusion (see 4.6.2) processes, and a tablet prepared from amelt-extruded solid dispersion (see 4.7). The dosing of the tablets andcapsules was followed with 20 ml of water whereas 10 mL of thesuspension formulations was dosed by gavage and followed by 10 mL ofwater to wash the gavage tube.

Blood samples were taken post-dose at 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7,12, 24 and 30 hours. The samples were centrifuged at 3000 rpm for 15minutes, the plasma removed into plain blood tubes and stored at −20° C.until analysis. Samples were analysed using a manual solid phaseextraction (Phenomenex Strata X, 30 mg) method followed by LC-MS usingthe conditions specified in Table 27 below

TABLE 27 Summary of LC-MS conditions for the determination of Compound 1in dog plasma Chromatographic conditions Apparatus Liquid chromatographwith MS/MS detector Column Waters Xterra Phenyl, 2.1 mm × 50 mm (3.5 μm)or equivalent Eluents Ammonium formate (1 mM, pH 3.0)/ Acetonitrile73:27 v/v) Flow rate 0.2 mL/min approx. Temperature 40° C. Wavelength276 nm Injection volume 5 μL Run time 4 min. Compound 1 2.7 min approx.retention time Mass spectrometer parameters Mode of operation Ion Spray(positive ion) (MS/MS) Voltage 4500 V approx. Temperature 450° C.approx. Ions monitored 435.3 to 281.2

6.2 Results

TABLE 28 Summary of pharmacokinetic data for Compound 1 formulationsBioavail- ability relative to copovidone solid AUC_((0-inf)) C_(p)maxdispersion Formulation (ng · hr ·mL⁻¹) (ng · mL⁻¹) capsule (%) ImmediateRelease tablet 7748 1225 19 (25% drug loading) Gelucire 44/14 (capsules,15649 2551 38 10% drug loading) Gelucire 44/14 (capsules, 10078 1654 2520% drug loading) Gelucire 44/14 (capsules, 7579 1174 18 40% drugloading) Microsuspension (1% drug 9327 1249 23 loading) Nanosuspension(1% drug 22746 3922 55 loading) Kleptose solid dispersion¹ 40373 7959 98(capsule; 20% drug loading, 1:3 drug:polymer ratio) PVP Soliddispersion¹ 35788 6810 87 (capsule; 20% drug loading, 1:3 drug:polymerratio) AQOAT solid dispersion¹ 35450 6840 86 (capsule; 20% drug loading,1:3 drug:polymer ratio) HPMC-606 solid 31739 6179 77 dispersion¹(capsule; 20% drug loading, 1:3 drug:polymer ratio) HP55S soliddispersion¹ 34687 6749 84 (capsule; 25% drug loading, 1:2 drug:polymerratio) Copovidone solid 41129 7707 100 dispersion² (capsule; 20% drugloading; 20:46 drug:polymer ratio) Copovidone solid 37745 7502 92dispersion (tablet; 25% drug loading; 30:70 drug:polymer ratio) ¹Blendedwith crospovidone disintegrant (100 mg/capsule) prior to filling²Blended with mannitol/Aerosil (99:1) (167 mg/capsule) prior to filling

See FIG. 30. Both C_(p)max and AUC from the polymer-based soliddispersions were significantly greater (P<0.05) than the immediaterelease tablet, Gelucire capsule and micro suspension/nano suspensionformulations.

1. An extrudate for use in an immediate release pharmaceuticalformulation, the extrudate formed during melt extrusion and comprisingan active agent in solid dispersion with a matrix polymer, wherein theactive agent is4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a salt or solvate thereof, and the matrix polymer is copovidone,wherein the ratio by weight of the active agent and matrix polymer is inthe range from 1:2 to 1:4.
 2. The extrudate as claimed in claim 1,wherein the active agent is in stable amorphous form.
 3. The extrudateas claimed in claim 2, wherein at least 90% by weight of the activeagent is in amorphous form.
 4. The extrudate as claimed in claim 1,wherein the copovidone is a co-polymer of 1-vinyl-2-pyrollidone andvinyl acetate in a ratio of 6:4 by mass.
 5. The extrudate as claimed inclaim 1, wherein the solid dispersion includes a surface-active agentand/or plasticiser.
 6. The extrudate as claimed in claim 5, wherein thesurface-active agent is selected from: sodium dodecyl sulphate (sodiumlauryl sulphate); docusate sodium; cetrimide; benzethonium chloride;cetylpyridinium chloride; lauric acid; polyoxyethylene alkyl ethers;polyoxyethylene sorbitan fatty acid esters polysorbates 20, 40, 60 and80; polyoxyethylene castor oil derivatives, e.g. Cremophor RH40™;polyoxyethylene stearates and poloxamers.
 7. A method of producing anextrudate according to claim 1, comprising: (i) mixing a suitable amountof4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof with a desiredamount of copovidone wherein the ratio by weight of the active agent tocopovidone is in the range of 1:2 to 1:4; (ii) increasing thetemperature of the mixture to produce a melt; and (iii) extruding themelt to produce a solid product.
 8. The method as claimed in claim 7,wherein in step (iii) the melt is extruded as rods or into one or moremoulds.
 9. The extrudate as claimed in claim 5, wherein the plasticizeris selected from acetyltributyl citrate, acetyltriethyl citrate, benzylbenzoate, chlorbutanol, dextrin, dibutyl phthalate, diethyl phthalate,dimethyl phthalate, glycerine, glycerine monostearate, mannitol, mineraloil, lanolin alcohols, palmitic acid, polyethylene glycol, polyvinylacetate phthalate, propylene glycol, 2-pyrrolidone, sorbitol, stearicacid, triacetin, tributyl citrate, triethanolamine and triethyl citrate.10. The extrudate as claimed in claim 1, wherein the solid dispersionfurther comprises a glidant.
 11. The extrudate as claimed in claim 10,wherein the glidant is colloidal silicon dioxide.
 12. The extrudate asclaimed in claim 1, wherein the extrudate is in the form of rods ormoulds.
 13. A method of producing an immediate-release pharmaceuticalformulation comprising: (i) mixing4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneor a pharmaceutically acceptable salt or solvate thereof withcopovidone, wherein the ratio by weight of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-oneto copovidone is in the range of from 1:2 to 1:4; (ii) increasing thetemperature of the mixture to produce a melt; (iii) extruding the meltto produce an extrudate according to claim 1; (iv) grinding theextrudate; (v) optionally mixing the ground extrudate with one or moreadditional excipients or ingredients; and (vi) tabletting into asuitable dosage form wherein the total concentration of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein the composition is in the range of from 20% by weight to 30% byweight and wherein the total amount of amount of4-[3-(4-cyclopropanecarbonyl-piperazine-1-carbonyl)-4-fluoro-benzyl]-2H-phthalazin-1-onein the composition is in the range from 25 mg to 400 mg.
 14. The methodaccording to claim 13, wherein the mixture formed in step (i) furthercomprises a glidant.